Journal of Biotechnology Research Center

Background: Salinization negatively affects plant growth and development, removing up to 1.5 million hectares of farmland from production annually. Objective: Evaluate the synergistic of Nano-fertilizers on yield traits of wheat Triticum aestivum below saline irrigation. Methodology: two factorial plot experiments were conducted during Autumn 2022 and 2023 in Diyala, IRAQ. The plants were irrigated every 18 days from planting with three NaCl irrigation levels: 0 (distilled water), 50, and 75 mM. A foliar application of nano fertilizer was sprayed after the third leaf appeared every 15 days, three times in total. Results: The salinity significantly decreased the yield component, biological yield, and harvest index, while the synergistic effects of (S), (Q), and (M) increased by 52.86%, 15.81%, and 18.84% in 2022 and 41.05%, 20.65% and 22.62 in 2023 growth season, respectively. Additionally, the most significant effects of the synergistic nano-fertilizer on catalase, peroxidase, and Proline under salinity were 23.84%, 70.00%, and 79.78% in 2022 and 24.50%, 73.33% and 74.36% in 2023, respectively. Conclusions: The application of nano-fertilizers enhanced yield parameters and increased the antioxidant enzymes as part of the plant's defense mechanism against oxidative stress. Over time, this response to developing salinity-tolerant wheat varieties with strong antioxidant defense systems.

Background: Breast cancer is a major public health problem, with a high
incidence among females worldwide, increasing with age and environmental
conditions. Hormonal biomarkers have gained attention for their potential
utility in the early detection of breast cancer. The study aimed to investigate the
effect of biomarkers of different hormone levels, including prolactin,
Testosterone, Cortisol, and Human Chorionic Gonadotropin (HCG) in the
diagnosis and early detection of breast cancer risk. Methodology: Blood and
saliva samples were collected from volunteer females, including 50 healthy
females (NB), 111 benign breasts (BB), and 20 malignant breasts (MB) samples
to determine hormone level biomarkers using enzyme-linked immunosorbent
assay (ELISA). Results: The results showed that hormonal biomarkers,
particularly prolactin, testosterone, and HCG, can serve as indicators for the
early detection of breast cancer in both serum and saliva. In contrast, cortisol
hormonal biomarkers did not have an effective association with early breast
cancer detection. Conclusion: The results of this study showed that the present
association between hormonal biomarkers, including prolactin, testosterone,
and HCG, could serve as biomarkers for the early detection of breast cancer. In
contrast, cortisol did not have an effective association with early breast cancer
detection.

Background: Pumpkin seeds are valuable source of good quality proteins from the amino acids that are made of them and can be used as functional food ingredients for their properties such as solubility, foam formation and stability. The aim of this study is to produce protein isolate and its enzymatic hydrolysates from local pumpkin seeds to study their properties.
Methodology: preparing defatted pumpkin seeds for extracting the protein isolate, and hydrolysate this protein by using Trypsin and pepsin enzymes separately and together by two ways. Determination of the amino acids and the degree of the hydrolysis was conducted, more over protein properties were determined such as solubility, emulsifying activity and stability index, foaming capacity and stability.
Results: Protein isolate was achieved from local peeled non soaked pumpkins seeds with protein percentage 53.15%. Enzymatic protein hydro-lysates were produced from this isolate at different times. Essential and non-essential amino acids were determined. Functional properties for protein isolate and protein hydro-lysates were studied, just like solubility in different pH. The higher emulsifying capacity was for the enzymatic hydro-lysate by using pepsin, while the protein isolate dissolved in pH=8 had higher emulsifying stability. The enzymatic hydro-lysate by using pepsin showed the highest foaming capacity and mixed enzymes (Ma) (pepsin+ trypsin) showed higher foaming capacity and stability.
Conclusion: Pumpkin seeds isolated protein has many nutritional values that may be used in preparation protein rich food for athletics and as a source of Nitrogen in microbial culturing media.

Background: Withania somnifera L. is a famous medicinal plant that has been utilized in Global folk medicine for a long time. The leaves and roots of the plant are rich in withanolide constituents, which are primarily responsible for their therapeutic effectiveness. Objective: This study focuses on the differences in the content of withanolide compounds and biomass in W. somnifera plants under different treatments with silver nanoparticles (Ag-NPs). Results: The leaves of the field plant (in vivo cultures) showed a rise in content of 27-OH-withanolide, withaferine-A, and withastramonolide. While the leaves and roots in the in vitro cultures revealed a high content of withanolide-B and withanone. The high content of withanolide-A was mainly observed in callus cultures. Moreover, the findings showed a considerable reduction in callus biomass across different concentrations of Ag-NPs (1, 2, and 3 mg/L). Conversely, there was a noteworthy rise in the production of withanolides. Specifically, withastramonolide and withanolide-A exhibited significant increases at 2 mg/L Ag-NPs, showing 121% and 15% increases, respectively, compared to the control. Additionally, several other withanolide compounds, including 27-OH-withanolide, withaferin-A, withanone, and withanolide-B, accumulated significantly at 3 mg/L Ag-NPs, with percentage increases ranging from 67% to 277%. Additionally, the study evaluated the DPPH scavenging activity in Ag-NPs-treated samples, showing a substantial 64.4% increase at 3 mg/L, which suggests the presence of elevated active components or improved antioxidants in this treatment. Conclusion: The findings provide insights into the variations in withanolide content under different culture systems and demonstrate the potential of Ag-NPs treatment to enhance withanolide yield and antioxidant activity in Withania plant cultures.

Background: As natural compounds, especially from plants, are considered a rich source of therapeutic importance, researchers have reported the benefits of using such products in treatments for diseases affecting humans and other organisms for many years. The study included the extraction of phenolic compounds from four genera of plants of the Compositae family growing wild in western Iraq, including: Achilliea fragrantissima, Herba-alba Artemisia (wormwood), Lactuca serriola (Wild lettuce ), and Silybum marianum. Methods: Hydro-alcoholic extraction was employed by the cold maceration method with 75% Ethanol. A general chemical analysis of the secondary metabolites in the crude ethanolic extract was conducted for each of the four plants separately. Thin layer chromatography (TLC) was used to estimate the types of phenolic compounds qualitatively, and the total phenols in each extracted residue were quantified using a colorimetric method with a standard curve plotted for the phenol standard (Gallic acid). The HPLC technique was also employed to identify the types and concentrations of phenolic compounds extracted from each plant residue. The effect of total phenolics extracted from each plant on cultures of the Leishmanial parasite was also studied in 24-hour in vitro experiments, using different concentrations of each plant's extract. The violet color was then read at the wavelength (620 nm) using an ELISA Reader device to obtain the inhibition rates, as the intensity of the color is an expression of the number of living cells. Results: The results showed that the plants contain phenols, flavonoids, tannins, glycosides, saponins, and alkaloids; however, the wormwood extract yielded a negative result for alkaloids. The results of thin-layer chromatography analysis showed that the phenolic extracts of the four plants contained, qualitatively, Pyrogallol, Cinnamic acid, gallic acid, and Hydroquinone in varying proportions. The extract also contained some types of flavonoids. Results for HPLC showed that the four plants are rich in these phenolic compounds. For quantitative total phenolic compound results, Artemisia, the wormwood plant, contained the highest percentage, 1464.72 mg% dry plant powder. In contrast, the lowest percentage was found in the Silybum plant, at 223.86 mg% dry plant powder. The activity against Leishmania growth inhibition rate showed that each plant extract exhibited a different mode of inhibiting the parasite cultures, with Silybum plant showing the best effects, and no significant difference was observed across all concentrations. These vary depending on the type of phenolic compounds and the flavonoids present in each plant, as well as their concentration. Conclusion: The four plant genera of the Compositae family growing wild in western Iraq, including: Achilliea fragrantissima, Herba-alba Artemisia (wormwood), Lactuca serriola (Wild lettuce), and Silybum marianum, are rich with phenolic compounds and different types of flavonoids which may help in parasite diminution, as these secondary active metabolites possess notable effects in free radical scavenging and antioxidant activity.

Background: Pyocins are narrow-spectrum bacteriocins that kill bacteria closely related to the producing strain and play a key role in colonization and competition in bacterial communities. This study aims to investigate the antibacterial and antibiofilm activity of pyocin.
Methodology: Twenty bacterial isolates of Pseudomonas aeruginosa were isolated from wounds and burns; the bacteriocin-producing isolates were detected using the cup assay method. Then, the bacteriocin was extracted from a highly producing strain, and this bacteriocin was tested against bacterial isolates other than the producing strain in both planktonic and biofilm form. The ability to form biofilm was tested using the microtiter plate method to isolate isolates that form strong biofilms. The antibiofilm activity of bacteriocin was also studied in this study using the same procedure as that used for testing biofilm..
Results: The results showed that all bacterial isolates possess the ability to produce bacteriocin against each other, but not against their own bacteriocin, because each isolate contains an immunity gene against its bacteriocin. The results of the biofilm study showed that six isolates produced a strong biofilm, while others produced a moderate biofilm. Other findings showed that bacteriocin exhibited high activity against biofilms formed by potent isolates, with the percentage of inhibition ranging from 48 to 67%. Conclusion: The bacteriocin can be used to inhibit biofilm-producing isolates of the same species, making it a potential alternative to antibiotics.

Background: Artemisia hera-alba, Achillea fragrantissima are plants naturally grown as wild herbs in western regions of Iraq, especially in Anbar lands belonging to Artemisia, which is one of the larger genera in the family Asteraceae, and primarily used in medicine against anthelminthic and antimalarial, besides its traditional use as antibacterial, antifungal, as well as hypoglycemic effect. The current study aimed to highlight the critical impact of the phenolic compounds, as these plants were rich in.
Methodology: An extraction method for Artemisia herba-alba (AR) and Achillea fragrantissima (A), involving the processing of aerial parts through cold maceration. Both plant extracts underwent phytochemical general investigations, including qualitative and quantitative estimations, as well as antioxidant assays, to elucidate their biological activities, specifically their immune-boosting and modulatory effects.
Results: The results indicated that the plant extract contained rich phenolic compounds, including Pyrogallol, Hydroquinone, and Cinnamic acid. Besides, the following flavonoids were present in both extracts: Rutin, Quercetin, Apigenin, Kaempferol, and Luteolin. The immune modulation effects indicated that both plant extracts showed a proliferative impact for normal human lymphocytes in higher doses, while cytotoxic effects appeared in low doses.
Conclusion: The toxic effects of all used concentrations of both plants affected the normal viability of natural killer cells, indicating that the dose might be adjusted to avoid any toxic effects that appear in these plant extracts at different medication states.

Background: Fungal species that parasitize crops cause significant losses annually as they affect the quality and quantity of crops. Infected crops are considered a poor product that negatively affects the local and international agricultural sector. Therefore, the rapid and accurate early detection of the pathogen is crucial for the crop and to avoid losses. A loop-mediated isothermal amplification (LAMP) assay is used to detect about 100 pg fragments of DNA genes per reaction, which accurately detects fungal infections. This assay is used after isothermal amplification to detect genes without the use of PCR. In addition, the analysis of the whole genome (AGE), a global method for determining micro genes, includes experimental practice analysis and bioinformatics testing. Recent tests to detect pathogenic fungi include isothermal amplification analysis, polymerase chain reaction (PCR) techniques, overlapping, biomagnetic, and quantum biometric methods. One of the modern methods is the bioinformatics approach, which involves comparing the entire gene to identify distinctive and specialized areas of the fungus and designing polymer interaction tests for the species and genus to be diagnosed.
Objective: The current study aims to survey accurate and rapid methods for diagnosing fungi that infect local crops, with the goal of controlling these infections, finding solutions, and preventing crop damage.
Conclusion: Due to the increase in pathogenic fungi that cause plant diseases, there is a need to dedicate efforts to following developments in modern biological and molecular techniques and to work on the diagnosis of new fungal species. The traditional detection of pathological fungi yields dubious and inaccurate results, which do not facilitate effective treatment against this pathogenic fungus. It has proven its efficiency in accurately and rapidly diagnosing fungal species and diseases that do not exhibit symptoms. Although many assays focus on polymer chain reaction, most recent studies tend to use quantitative polymer chain reaction on a vast scale for quantitative measurement and differentiation between the factors causing fungal pests on plants, especially when the sample quantity is too small to detect, where the Lamb technique has succeeded in discovering many fungal species such as Alternaria spp., Fusarium spp., Puccinia spp., Colletotrichum spp., and Foma spp.. By amplifying the genomes of samples, modern genetic techniques have also succeeded in identifying fungal species and comparing them with the dolphin data without prior knowledge.

Background: TGF-β is the main regulator of immune responses and plays an important role in the development of autoimmune diseases in humans, and is strongly implicated in autoimmune thyroid diseases. Objective: This study aimed to evaluate the immunological and gene expression of TGF-β and their relationship to recurrent miscarriages
Methodology: The study was conducted in Iraq in Salahuddin province, during which 90 samples were collected from women,70 of whom were infected with AITD with positive TPOAb. They were compared with 20 healthy women.
Results: During this study, highly significant differences in TGF-β gene expression were observed at p ≤ 0.001 between patients with AITD and healthy individuals. The means were (1.739±0.214) and (1.208±0.347), with highly significant differences in the immunological TGF-β levels at p≤0.001 between pregnant patients and patients with RM, and the means were (1.7511±0.4955) and (1.0145±0.1960). It was also found that significant differences in the level of TH17 at p≤0.001 existed between pregnant patients and those with RM; the mean was (1.0111±0.2284) and (1.5298±0.2922), respectively.
Conclusion: Through this study, it was found that the presence of the TGF-β gene and its high levels maintain pregnancy and work to suppress the cytokines secreted by Th17, which stimulate miscarriage. Additionally, the high level of the gene enhances its immune response, while its low level activates Th17 cells and increases the secretion of inflammatory cytokines.

Background: Probiotics refer to live microorganisms that promote the organism's well-being when consumed in sufficient quantities. Lactobacillus acidophilus, a well-established probiotic, is often suggested for its positive impact on health.
Objective: To investigate the spread of antibiotic resistance and the antibacterial effectiveness of probiotic bacteria against pathogenic bacteria obtained from skin infections.
Methodology: A total of 40 samples were collected from patients with skin infections in August and October 2017 from two hospitals in Baghdad. The samples originated from patients of various ages and genders. Following standard morphological and biochemical characterization, 69 isolates were identified as Enterobacter cloacae (n=12, 17.4%), Klebsiella pneumoniae (n=1, 1.4%), Proteus mirabilis (n=1, 1.4%), Pseudomonas aeruginosa (n=15, 21.7%), and Staphylococcus aureus (n=21, 30.4%). Eleven isolates (15.9%) belonged to Staphylococcus epidermidis, with the remaining isolates distributed across Bacillus spp. (n=6, 8.6%), Pseudomonas stutzeri (n=1, 1.4%), and Enterobacter aerogenes (n=1, 1.4%). Antibiotic susceptibility testing using nine antibiotics identified 18 isolates resistant to Gentamicin, Cefotaxime, Amikacin, Ceftriaxone, Tobramycin, and Amoxicillin-clavulanic acid. L. acidophilus, a potential probiotic, was cultured and evaluated for its inhibitory activity against the isolated skin infection bacteria.
Results: The results upon evaluating the inhibitory effect of the probiotic L. acidophilus against bacteria causing skin infections demonstrated a broad-spectrum inhibitory impact at all tested concentrations against the isolated skin infection pathogens.
Conclusion: L. acidophilus, a probiotic bacterium, demonstrated inhibitory activity against skin infection-causing bacteria.

Background: The southern marshes of Iraq are a variety of aquatic environments; these include deep and shallow perennial, seasonal, fresh, and salt marshes, most of which are open water and dense aquatic vegetation.
Methodology: The study was conducted on the Hammer Marsh in the Al-Fhood sub-district in Dhi-Qar governorate. It is a group of marshes located south of Al-Shatrah, one of the permanent marshes; the length is about 50 km, and the study includes 9 stations.
Results: The study showed the dominance of (Phragmites australis) and (Typha domingensis) plants in wetlands, it was found in all study stations. The clear waters in the deeper areas of the marsh have provided support for the submerged and diverse aquatic plant groups, including (Ceratophyllum demersum) that appeared in four stations (F1, F2, F3, F5b) other scattered plant groups, and several existing swamp weeds appeared, including the (Myriphyllum vulgar) and (Potamogeton pectiatus). As for the phosphate values, they were higher than the permissible limits in most of the stations recorded (0.752mg/L) in station F5b, and the nitrate values were recorded at high values in marsh water, higher than the internationally permissible limits (1) that it recorded (1.1 mg/L) in station F2. The growth of aquatic plants appears dense in the Hammer marsh in Al-fhood, such as (Phragmites australis), (Typha domingensis), (Ceratophyllum demersum), (Myriophyllum), and (Potamogeton pectiatus), where it grew in the stations (F1, F2, F3, F5b). The phytonutrients provide phosphate and Nitrogen, and the ones coming from human waste, mainly in the areas surrounding the marsh, stimulate the growth of aquatic plants, which later decompose to produce unacceptable odors and add BOD to the water.
Conclusion: There is a difference in the ability of aquatic plants and weeds to absorb nutrients in the marsh water.

Background: Osteoporosis is a degenerative, progressive bone condition that causes widespread fragility and an increased risk of fractures. This study was designed to determine some levels of Immunological parameters, such as Interleukin (IL) IL-17, IL-1, IL-15, IL-6, Hematological parameters (Hb and WBC), and some biochemical parameters (Vit D3) in patients with osteoporosis and compare their parameter levels with the control group. Methodology: This cross-sectional study was conducted among the general population who attended Ramadi Teaching Hospital in Ramadi city from the 1st of November 2023 to the 1st of January 2024. The total number of blood samples were 80 samples which are included 50 samples for patients with osteoporosis with ages ranging from 16 to 50 years and 30 samples for healthy individuals as a control group . Results: There was significant deference in all parameters with p-value (<0.05), the means levels of IL-1, IL-17, IL-15, IL-6 and Vit D3 respectively were: (8.467pg\mL), (89.32pg\mL), (83.84pg\mL), (60.34pg\mL), (4.667pg\mL); As well as the concentration of Hb, and mean number of WBC were: Hb(9.16g/dL) and\ WBC(14.19*103 c/ mm3) respectively. Conclusion: We conclude that patients suffering from osteoporosis showed a significant increase in the levels of IL-1, IL-17, IL-15, IL-6, and WBC compared with controls. However, there was a significant decrease in vitamin D3 and Hb in patients compared to controls.

Background: Artificial sweeteners are widely consumed as sugar substitutes in various food products, as they are promoted for their low-calorie content, which helps manage blood sugar levels and aids in weight control.
Methodology: This study examines the adverse effects of artificial sweeteners on specific physiological parameters in albino rats. We used twenty-five male albino rats weighing 180-250 g. The animals were divided into five groups (control, sucrose, stevia, stevia leaves, and saccharin), with five rats in each group. The duration of the experiment lasted 45 days, and the sweeteners were dissolved individually in water. Results: Rats showed a preference for water consumption with sucrose and saccharine, but consumed less with stevia leaves. The blood was collected for serum glucose, Alanine Aminotransferase (ALT), Aspartate Aminotransferase (AST), blood urea, serum creatinine, lipid profile, and C-reactive protein (CRP) measurement. Results showed a significant increase in glucose levels (mg/dL) at 176±17%, 119±10%, and 132±13%, respectively. Urea mg/dL(33±3%, 28±2 % and 31±1%) s. creatinine mg/dL(0.8±0.2%, 0.5±0.1% and 0.9±0.1%), GPT U/L (45±4%, 33±3 % and 41±3%) GOT U/L (128 ±11 %, 70±9 % and 129 ±12 %) cholesterol mg/dL(196 ±20 %, 162 ±12 % and 209 ±18 %) triglyceride mg/dL(178 ±17 %, 119 ±11 % and 187 ±16 %), HDL mg/dL(25 ±2 , 46 ±5 % and 36 ±3 %) and VLDL mg/dL(35 ±3 %, 23 ± 2 % and 22 ±1 %) of (sucrose, stevia and saccharin) groups when we compared them with the control group. It was found that dietary supplementation with stevia leaves didn’t affect blood glucose107 ±15 %, b.urea 24 ±5%, s.creatnine 0.4 ±0.04%, GOT 64±6 %, cholesterol 153 ±15 %, triglyceride117 ± 2% , HDL46±5 and VLDL 37±3 %. Except for GPT, there was a significantly elevated 33±3 % when we compared with the control group. Also, there were no significant changes in alkaline phosphatase levels in all five groups. As for CRP, there was no significant difference between the five groups; all groups were negative. Conclusion: It can be concluded that sweeteners must be used carefully because they have potential side effects due to their action on the Liver, kidneys, and lipid profile, as they cause significant changes in blood biochemical tests. It’s vital to create high consciousness for consumers regarding its effects, especially for children .